The HNRNPD Knockout HEK293T Polyclonal Cells product provides a CRISPR/Cas9-mediated gene disruption model targeting the HNRNPD gene in a polyclonal HEK293T background. These loss-of-function cells enable investigation of HNRNPD-dependent post-transcriptional regulation, avoiding clonal biases inherent to monoclonal isolates. HNRNPD encodes AUF1, an AU-rich element (ARE)-binding protein central to mRNA decay and translation control, making this population suitable for diverse molecular analyses.
HEK293T cells are human embryonic kidney epithelial cells that stably express the SV40 large T antigen, conferring high transfectability and permitting episomal replication of plasmids carrying the SV40 origin. Widely employed for transient gene expression and viral vector production, this line retains epithelial characteristics that provide a relevant context for studying signaling and gene regulation. Its robust growth and ease of manipulation make HEK293T an optimal host for CRISPR-based knockout studies.
HNRNPD (AUF1) binds ARE motifs in the 3′ UTRs of labile mRNAs, directing them toward accelerated decay or stabilization depending on cellular context and post-translational modifications. It integrates signals from upstream regulators including TNF-??, MAPK, and NF-??B, and controls key targets such as c-fos, c-myc, cyclin D1, TNF-??, and IL-6. Mechanistically, HNRNPD interacts with other ARE-binding proteins like HuR and tristetraprolin (TTP) and recruits the exosome complex and PARN deadenylase to execute rapid mRNA turnover. This network forms a dynamic hub that fine-tunes gene expression in processes like cell proliferation and inflammation.
In HEK293T cells, HNRNPD disruption provides a tractable system to analyze ARE-mediated mRNA decay. The epithelial origin and high transfectability allow detailed dissection of how HNRNPD modulates oncogene and cytokine expression. Loss of HNRNPD can differentially alter the stability of target transcripts, thereby affecting cell cycle progression and inflammatory responses. This model offers a physiologically relevant platform for linking post-transcriptional regulation to cellular phenotypes in cancer and immune dysregulation.
Applications include actinomycin D chase assays for mRNA half-life measurement, RNA immunoprecipitation to probe protein-RNA complexes, and ARE-luciferase reporter assays for regulatory element activity. Western blotting and RT-qPCR validate downstream target expression, while co-immunoprecipitation and immunofluorescence identify interacting partners and subcellular localization. This knockout tool supports research in oncology, RNA biology, and inflammation. For further technical details, please contact Ascent Research.
